OFFGEL electrophoresis and tandem mass spectrometry approach compared with DNA-based PCR method for authentication of meat species from raw and cooked ground meat mixtures containing cattle meat, water buffalo meat and sheep meat

Liquid Chromatography–mass Spectrometry Bottom‐up Proteomic Methods in Animal Species Analysis of Processed Meat for Food Authentication and the Detection of Adulterations

“…Several different heat treatments were employed to achieve a high degree of protein denaturation: cooking in an oven at 180°C for 1 hr (Sentandreu et al, 2010), boiling at 100°C for 30 min (Sarah et al, 2016), autoclaving at 121°C for 20 min (Claydon et al, 2015;Sarah et al, 2016), heating in an oven at 190°C until the meat reached a core temperature of 99°C Montowska & Fornal, 2017, cooking at 350°C until the meat reached 71°C internal temperature (Ruiz Orduna et al, 2017), sealing with bags and cooking in a boiled water bath at 100°C for 30 min (Naveena et al, 2017), grilling at 150°C and 180°C both for 10 min (Wang et al, 2018), heating in oven at 200°C until meat reached a core temperature of 100°C (Von Bargen et al, 2014), frying in a Teflon-coated pan until a brown crust is formed (Von Bargen et al, 2014), cooking at 90-100°C for 2 hr and autoclaving to 120°C (cycle time 90 min) (Prandi et al, 2019), frying in boiling oil until golden brown (Li et al, 2018), baking at 200°C for 30 min (Li et al, 2018), simmering for 20 min and then autoclaving at 120°C for 15 min (Claydon et al, 2015). Some authors had also proposed the washing of samples after heat treatment processing Montowska & Fornal, 2017.…”

“…Von Bargen et al (2014) also proposed preparing the meat mixture by mixing desalted peptide solutions of the relevant species. Two-component mixtures (beef with pork [Von Bargen et al, 2014;Ruiz Orduna et al, 2017], beef with horse [Von Bargen et al, 2014;Ruiz Orduna et al, 2017] sheep with buffalo [Naveena et al, 2017], beef with buffalo [Naveena et al, 2017]), three-component mixtures (chicken with turkey and pork [Montowska & Fornal, 2017, beef with lamb and duck [Li et al, 2018]), four-component mixtures (horse with beef, pork, and chicken [Claydon et al, 2015], pork with beef, chicken, and sheep [Pan et al, 2018], beef and pork with sheep and goat [Prandi et al, 2019], lamb with pork, chicken, and beef [Li et al, 2018]) and six-components mixtures (beef and pork with donkey, buffalo, horse, and red deer [Prandi et al, 2019], beef and pork with duck, rabbit, chicken, and turkey [Prandi et al, 2019]) were most often prepared, where the content of one meat spiked with another was in the range from 0.5% to 50%.…”

“…In one approach, raw meat is thermally processed before it is homogenized, and then homogenates are subjected to an extraction procedure (Li et al, 2018). In the second approach, raw meat is homogenized, the homogenates are sealed in bags and cooked in a water bath and then cooked subsamples are subjected to an extraction assay (Naveena et al, 2017). Homogenization procedures include cutting meat into slices, mincing, or grinding (Li et al, 2018).…”

“…One of the first developed techniques for meat identification were electrophoretic based methods such as sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) (Naveena et al, 2017) and one-, two-dimensional gel electrophoresis (1-DE, 2-DE) (Kim et al, 2017). As reported by von Bargen's group (Von Bargen et al, 2013), a vast number of SDS-PAGE methods were available at the end of the 1980s.…”

“…The OFFGEL enrichment fractionation step and MS detection by LC-MS/MS were implemented to detect as low as 0.5% (w/w) contaminating chicken in pork cooked meat (Sentandreu et al, 2010). Naveena et al (2017) have demonstrated the suitability of an OFFGEL electrophoresis-mass spectrometry based proteomic approach for identification of species-specific peptides derived from myosin light chain 1 and myosin light chain 2 for detection of contaminating buffalo meat in sheep meat as low as 0.5% (w/w) in both raw and processed meat.…”